ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2008, Vol. 39 ›› Issue (11): 1548-1553.doi:

• 预防兽医 • Previous Articles     Next Articles

Development and Application of the Recombinant ELISA Based on the Assembling and Expression of VP7 Gene of African Sickness Virus

GAO Zhi-qiang，ZHANG He-xiao*，LAI Ping-an，GU Qiang，PU Jing，
WANG Lin，QIAO Cai-xia，WU Dan，BAI Ya-duo，ZHANG Wei   

  1. Animal Quarantine Center of Beijing EntryExit Inspection and Quarantine Bureau, Beijing 101113, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-11-24 Published:2008-11-24

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Abstract: A DNA fragment, which encodes the most linear epitopes of VP7 of African sickness virus(AHSV), was assembled artificially, then cloned into pET-30a. Target protein expressed at very high level as inclusion body after the recombinant pET-30a-VP7 plasmid was transformed into BL21(DE3), and induced with 1.0 mmol/L IPTG. The indirect ELISA for detecting AHSV VP7 protein antibody was established after the reaction activity of the recombinant protein was determined by Dot-ELISA and ELISA. The results obtained indicated that the optimum antigen concentration was 0.25 μg/mL, and optimum serum dilution was 1∶40. The cut-off value was determined as 0.25. 184 serum samples were detected by this method and commercial ELISA kit and the results were all negative.